This invention relates to the field of biology. In a particular embodiment, it relates to compositions useful to monitor or elicit an immune response to selected tumor-associated antigens.
I. Background of the Invention
II. Summary of the Invention
III. Brief Description of the Figures
IV. Detailed Description of the Invention
A. Definitions
B. Stimulation of CTL and HTL responses
C. Binding Affinity of Peptide Epitopes for HLA Molecules
D. Peptide Epitope Binding Motifs and Supermotifs
1. HLA-A2 supermotif
2. HLA-A2.1 motif
3. HLA Class II Motifs and PADRE(trademark)
E. Enhancing Population Coverage of the Vaccine
F. Immune Response-Stimulating Peptide Epitope Analogs
G. Preparation of Peptide Epitopes
H. Assays to Detect T-Cell Responses
I. Use of Peptide Epitopes for Evaluating Immune Responses
J. Vaccine Compositions
1. Minigene Vaccines
2. Combinations of CTL Peptides with Helper Peptides
3. Combinations of CTL Peptides with T Cell Priming Materials
4. Vaccine Compositions Comprising Dendritic Cells Pulsed with CTL and/or HTL Epitopes
K. Administration of Vaccines for Therapeutic or Prophylactic Purposes
L. Kits
V. Examples
VI. Claims
VII. Abstract
The field of immunotherapy is yielding new approaches for the treatment of cancer, including the development of improved cancer vaccines (Krul, K. G., Decision Resources, 10.1-10.25 (1998)). While vaccines provide a mechanism of directing immune responses towards the tumor cells, there are a number of mechanisms by which tumor cells circumvent immunological processes (Pardoll, D. M., Nature Medicine (Vaccine Supplement), 4:525-531 (1998)). Recent advances indicate that the efficacy of peptide vaccines may be increased when combined with approaches which enhance the stimulation of immune responses, such as the use of Interleukin-2 or autologous dendritic cells (DC) (Abbas et al., eds., Cellular and Molecular Immunology, 3rd Edition, W. B. Saunders Company, pub. (1997)).
In a Phase I study, Murphy, et al., demonstrated that Human Leukocyte Antigen (HLA)-A2-binding peptides corresponding to sequences present in prostate specific antigen (PSA) stimulated specific cytotoxic T-cell lymphocyte (CTL) responses in patients with prostate cancer (Murphy et al., The Prostate 29:371-380 (1996)). Recently, Rosenberg, et al., evaluated the safety and mechanism of action of a synthetic HLA-A2 binding peptide derived from the melanoma associated antigen, gp100, as a cancer vaccine to treat patients with metastatic melanoma (Rosenberg et al., Nature Med., 4:321-327 (1998)). Based on immunological assays, 91% of patients were successfully immunized with the synthetic peptide. In addition, 42% (13/31) of patients who received the peptide vaccine in combination with IL-2 treatment, demonstrated objective cancer responses. Finally, Nestle, et al., reported the vaccination of 16 melanoma patients with peptide- or tumor lysate-pulsed DC (Nestle et al., Nature Med 4:328-332 (1998)). Peptide-pulsed DC induced immune responses in (11/12) patients immunized with a vaccine comprised of 1-2 peptides. Objective responses were evident in 5/16 (3 peptide-pulsed, 2 tumor-lysate pulsed) evaluated patients in this study. These Phase I safety studies provided evidence that HLA-A2 binding peptides of known tumor-associated antigens demonstrate the expected mechanism of action. These vaccines were generally safe and well tolerated. Vaccine molecules related to four cancer antigens, CEA, HER2/neu, MAGE2, and, MAGE3 have been disclosed. (Kawashima et al., Human Immunology, 59:1-14 (1998)).
Preclinical studies have shown that vaccine-pulsed DC mediate anti-tumor effects through the stimulation of antigen-specific CTL (Mandelboim et al., Nature Med., 1: 1179-1183 (1995); Celluzzi et al., J Exp Med 183:283-287 (1996); Zitvogel et al., J Exp Med 183:87-97 (1996); Mayordomo et al., Nature Med 1:1297-1302 (1995)). CTL directly lyse tumor cells and also secrete an array of cytokines such as interferon gamma (IFNxcex3), tumor necrosis factor (TNF) and granulocyte-macrophage colony stimulating factor (GM-CSF), that further amplify the immune reactivity against the tumor cells. CTL recognize tumor associated antigens (TAA) in the form of a complex composed of 8-11 amino acid residue peptide epitopes, bound to Major Histocompatibility Complex (MHC) molecules (Schwartz, B. D., The human major histocompatibility complex HLA in basic and clinical immunology Stites et al., eds., Lange Medical Publication: Los Altos, pp. 52-64, 4th ed.). Peptide epitopes are generated through intracellular processing of 3proteins. The processed peptides bind to newly synthesized MHC molecules and the epitope-MHC complexes are expressed on the cell surface. These epitope-MHC complexes are recognized by the T cell receptor of the CTL. This recognition event is required for the activation of CTL as well as induction of the effector functions such as lysis of the target tumor cell.
MHC molecules are highly polymorphic proteins that regulate T cell responses (Schwartz, B. D., The human major histocompatibility complex HLA in basic and clinical immunology Stites et al., eds., Lange Medical Publication: Los Altos, pp. 52-64, 4th ed.). The species-specific MHC homologues that display CTL epitopes in humans are termed HLA. HLA class I molecules can be divided into several families or xe2x80x9csupertypesxe2x80x9d based upon their ability to bind similar repertoires of peptides. Vaccines which bind to HLA supertypes such as A2, A3, and B7, will afford broad, non-ethnically biased population coverage. As seen in Table 11, population coverage is 84-90% for various ethnicities, with an average coverage of the sample ethnicities at 87%.
Various approaches have, or are, being employed as cancer vaccines. Table 1 overviews the major cancer vaccine approaches and the various advantages and disadvantages of each.
Currently there are a number of unmet needs in the area of cancer treatment. This is evidenced by the side effects associated with existing therapies employed for cancer treatment and the fact that less than 50% of patients are cured by current therapies. Therefore, an opportunity exists for a product with the ability to either increase response rates, duration of response, overall survival, disease free survival or quality of life.
Disclosed herein is a composition comprising all eight isolated epitopes: YLSGANLNV (SEQ ID NO:1), IMIGVLVGV (SEQ ID NO:2), KLBPVQLWV (SEQ ID NO:3), SMPPPGTRV (SEQ ID NO:4), KVAELVHFL (SEQ ID NO:5), YLQLVFGIEV (SEQ ID NO:6), RLLQETELV (SEQ IS NO:7), and VVLGVVFGI (SEQ ID NO:8). The composition can further comprise an antigen presenting cell, whereby at least one epitope is bound to an HLA molecule on the antigen presenting cell, such that a T lymphocyte receptor can bind to a complex of the HLA molecule and the epitope. The antigen presenting cell can be a dendritic cell. The composition can comprise each epitope connected to another epitope by peptide bonds. An amino acid linker can be a component of the composition, wherein at least two of the epitopes are connected to each other by peptide bonds. The composition can comprise a CTL or HTL epitope; the HTL epitope can be a pan-DR binding molecule. The composition can comprise a liposome, wherein the epitopes are on or within the liposome. A lipid can be attached to one of the epitopes.
In another embodiment, a composition can comprise at least one peptide, the peptide comprising an isolated, prepared epitope consisting of a sequence selected from the group consisting of: VLYGPDAPTV (SEQ ID NO:9), YLSGANLNV (SEQ ID NO:1), ATVGIMIGV (SEQ ID NO:10), LLPENNVLSPV (SEQ ID NO:11), KLCPVQLWV (SEQ ID NO:12), KLBPVQLWV (SEQ ID NO:3), SLPPPGTRV (SEQ ID NO:13), SMPPPGTRV (SEQ ID NO:4), KLFGSLAFV (SEQ ID NO:14), KVFGSLAFV (SEQ ID NO:15), VMAGVGSPYV (SEQ ID NO:16), ALCRWGLLL (SEQ ID NO:17), FLWGPPALV (SEQ ID NO:18), HLYQGCQVV (SEQ ID NO:19), ILHNGAYSL (SEQ ID NO:20), IMIGVLVGV (SEQ ID NO:2), KIFGSLAFL (SEQ ID NO:21), KVAELVHFL (SEQ ID NO:5), LLTFWNPPV (SEQ ID NO:22), LVFGIELMEV (SEQ ID NO:23), QLVFGIELMEV (SEQ ID NO:24), RLLQETELV (SEQ ID NO:7), VVLGVVFGI (SEQ ID NO:8), YLQLVFGIEV (SEQ ID NO:6), and YMIMVKCWMI (SEQ ID NO:25). The epitope can be joined to an amino acid linker. The epitope can be admixed or joined to a CTL epitope. The epitope can be admixed or joined to an HTL epitope, which can be a pan-DR binding molecule. The composition can further comprise a liposome, wherein the epitope is on or within the liposome. The epitope can be joined to a lipid. An epitope of the composition can br present as a heteropolymer or a homoplymer. The composition can comprise an HLA heavy chain, wherein the epitope contacts the HLA heavy chain. The HLA heavy chain can contact xcex22-microglobulin, biotin or, streptavidin. The HLA heavy chain can contact xcex22-microglobulin and biotin, and, the biotin can contact streptavidin, whereby a tetrameric complex is formed. The composition can further comprise an antigen presenting cell, wherein the epitope is on or within the antigen presenting cell. When the epitope is bound to an HLA molecule on the antigen presenting cell, whereby, when an A2-restricted cytotoxic lymphocyte (CTL) is present, a receptor of the CTL binds to a complex of the HLA molecule and the epitope. The antigen presenting cell can be a dendritic cell.
A further embodiment of the invention comprises a peptide comprising less than 50 contiguous amino acids that have 100% identity with a native peptide sequence of HER2/neu, MAGE2, MAGE3, p53, or CEA; the peptide further comprising an epitope selected from the group consisting of: YLSGANLNV (SEQ ID NO:1), KLBPVQLWV (SEQ ID NO:3), and SMPPPGTRV (SEQ ID NO:4). The peptide can comprise epitopes from two sequences selected from the group of native peptide sequences consisting of: HER2/neu, MAGE2, MAGE3, p53, and CEA. The peptide can comprise two or three epitopes selected from the group consisting of: YLSGANLNV (SEQ ID NO:1), KLBPVQLWV (SEQ ID NO:3), and SMPPPGTRV (SEQ ID NO:4). The peptide can comprise an additional epitope. The additional epitope can be selected from the group consisting of: ALCRWGLLL (SEQ ID NO:17), FLWGPRALV (SEQ ID NO:18), HLYQGCQVV (SEQ ID NO:19), ILHNGAYSL (SEQ ID NO:20), IMIGVLVGV (SEQ ID NO:2), KIFGSLAFL (SEQ ID NO:21), KVAELVHFL (SEQ ID NO:5), LLTFWNPPV (SEQ ID NO:22), LVFGIELMEV (SEQ ID NO:23), QLVFGIE (SEQ ID NO:24), RLLQETELV (SEQ ID NO:7), VVLGVVFGI (SEQ ID NO:8), YLQLVFGIEV (SEQ ID NO:6), and YMIMVKCWMI (SEQ ID NO:25). The additional epitope can be selected from the group consisting of: IMIGVLVGV (SEQ ID NO:2), KVAELVHFL (SEQ ID NO:5), YLQLVFGIEV (SEQ ID NO:6), RLLQETELV (SEQ ID NO:7), and VVLGVVFGI (SEQ ID NO:8). The additional epitope can be selected from the group consisting of: VLYGPDAPTV (SEQ ID NO:9), YLSGANLNV (SEQ ID NO:1), ATVGIMIGV (SEQ ID NO:10), LLPENNVLSPV (SEQ ID NO:11), KLCPVQLWV (SEQ ID NO:12), KLBPVQLWV (SEQ ID NO:3), SLPPPGTRV (SEQ ID NO:13), SMPPPGTRV (SEQ ID NO:4), KLFGSLAFV (SEQ ID NO:14), KVFGSLAFV (SEQ ID NO:15), and VMAGVGSPYV (SEQ ID NO:16). The additional epitope can be a PanDR binding molecule. The peptide can be a heteropolymer or a homopolymer.
An alternative embodiment of the invention is a composition comprising one or more peptides, and further comprising at least six epitopes selected from the group consisting of: YLSGANLNV (SEQ ID NO:1), IMIGVLVGV (SEQ ID NO:2), KLBPVQLWV (SEQ ID NO:3), SMPPPGTRV (SEQ ID NO:4), KVAELVHFL (SEQ ID NO:5), YLQLVFGIEV (SEQ ID NO:6), RLLQETELV (SEQ ID NO:7), and VVLGVVFGI (SEQ ID NO:8), wherein each of said one or more peptides comprise less than 50 contiguous amino acids that have 100% identity with a native peptide sequence. The composition can comprise one peptide that comprises the at least six epitopes. The composition can comprise an epitope from p53. The composition can comprise seven or eight epitopes selected from the group consisting of: YLSGANLNV (SEQ ID NO:1), IMIGVLVGV (SEQ ID NO:2), KLBPVQLWV (SEQ ID NO:3), SMPPPGTRV (SEQ ID NO:4), KVAELVHFL (SEQ ID NO:5), YLQLVFGIEV (SEQ ID NO:6), RLLQETELV (SEQ ID NO:7), and VVLGVVFGI (SEQ ID NO:8). The peptide of the composition can be a heteropolymer or a homopolymer. The composition can further comprise an additional epitope. The additional epitope can be derived from a tumor associated antigen, or be a PanDR binding molecule.
A further embodiment of the invention comprises a vaccine composition comprising: a unit dose of a peptide that comprises less than 50 contiguous amino acids that have 100% identity with a native peptide sequence of CEA, HER2/neu, MAGE2, MAGE3, or p53; the peptide comprising an epitope selected from the group consisting of: VLYGPDAPTV (SEQ ID NO:9), YLSGANLNV (SEQ ID NO:1), ATVGIMIGV (SEQ ID NO:10), LLPENNVLSPV (SEQ ID NO:11), KLCPVQLWV (SEQ ID NO:12), KLBPVQLWV (SEQ ID NO:3), SLPPPGTRV (SEQ ID NO:13), SMPPPGTRV (SEQ ID NO:4), KLFGSLAFV (SEQ ID NO:14), KVFGSLAFV (SEQ ID NO:15), VMAGVGSPYV (SEQ ID NO:16), ALCRWGLLL (SEQ ID NO:17), FLWGPRALV (SEQ ID NO:18), HLYQGCQVV (SEQ ID NO:19), ILHNGAYSL (SEQ ID NO:20), IMIGVLVGV (SEQ ID NO:2), KIFGSLAFL (SEQ ID NO:21), KVAELVHFL (SEQ ID NO:5), LLTFWNPPV (SEQ ID NO:22), LVFGIELMEV (SEQ ID NO:23), QLVFGIELMEV (SEQ ID NO:24), RLLQETELV (SEQ ID NO:7), VVLGVVFGI (SEQ ID NO:8), YLQLVFGIEV (SEQ ID NO:6), and YMIMVKCWMI (SEQ ID NO:25); and; a pharmaceutical excipient. The composition can further comprise an additional epitope. The additional epitope can be selected from the group consisting of: ALCRWGLLL (SEQ ID NO:17), FLWGPRALV (SEQ ID NO:18), HLYQGCQVV (SEQ ID NO:19), ILHNGAYSL (SEQ ID NO:20), IMIGVLVGV (SEQ ID NO:2), KIFGSLAFL (SEQ ID NO:21), KVAELVHFL (SEQ ID NO:5), LLTFWNPPV (SEQ ID NO:22), LVFGIELMEV (SEQ ID NO:23), QLVFGIELMEV (SEQ ID NO:24), RLLQETELV (SEQ ID NO:7), VVLGVVFGI (SEQ ID NO:8), YLQLVFGIEV (SEQ ID NO:6), and YMIMVKCWMI (SEQ ID NO:25). The additional epitope can be a PanDR binding molecule. The pharmnaceutical excipient can comprise an adjuvant. The vaccine composition can flirther comprise an antigen presenting cell, whereby when the epitope is bound to an HLA molecule on the antigen presenting cell, and when an A2 supertype-restricted cytotoxic T lymphocyte (CTL) is present, a receptor of the CTL binds to a complex of the HLA molecule and the epitope. The antigen presenting cell can be a dendritic cell. The vaccine composition can further comprise a liposome, wherein the at least one epitope is on or within the liposome.
Disclosed herein is a composition comprising one or more peptides, and further comprising at least one epitope selected from Table 6, wherein each of said one or more peptides comprise less than 250, 100, 75 or 50 contiguous amino acids that have 100% identity with a native peptide sequence. The native peptide sequence can be from an antigen selected from the group consisting of CEA, HER2/neu, MAGE2, MAGE3, p53 and a prostate tumor associated antigen. The at least one epitope can be YLSGANLNV (SEQ ID NO:1), IMIGVLVGV (SEQ ID NO:2), KLBPVQLWV (SEQ ID NO:3), SMPPPGTRV (SEQ ID NO:4), KVAELVHFL (SEQ ID NO:5), YLQLVFGIEV (SEQ ID NO:6), RLLQETELV (SEQ ID NO:7), and VVLGVVFGI (SEQ ID NO:8). The composition can further comprise a pharmaceutical excipient, such as an adjuvant.
An alternative embodiment comprises, a composition comprising one or more peptides, and further comprising at least two epitopes selected from Table 6, wherein each of said one or more peptides comprise less than 250, 100, 75 or 50 amino acids that have 100% identity with a native peptide sequence. A native peptide sequence can be CEA, HER2/neu, MAGE2, MAGE3, or p53. The composition comprises epitopes from at least two antigens selected from the group consisting of antigens: CEA, HER2/neu, MAGE2, MAGE3, and p53. The composition can comprise at least two, three, four, five, six, seven, eight, nine, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or more epitopes selected from Table 6. A multiepitope peptide can be a heteropolymer or a homopolymer. The composition can comprise at least one epitope selected from Table 6 which binds to a member of an HLA-A2 supertype at an IC50 equal to or less than 500 nM, at least one epitope selected from Table 6 which binds to at least two members of the HLA-A2 supertype at an IC50 equal to or less than 500 nM, at least one epitope selected from Table 6 which binds to at least three members of the HLA-A2 supertype at an IC50 at equal to or less than 500 nM, at least one epitope selected from Table 6 which binds to a member of the HLA-A2 supertype at an IC50 at equal to or less than 200 nM, at least one epitope selected from Table 6 which binds to a member of the HLA-A2 supertype at an IC50 at equal to or less than 50 nM, or, at least one epitope selected from Table 6 which exhibits tumor cell recognition when used in a CTL assay to elicit a CTL response. The composition can comprise a pharmaceutical excipient, such as an adjuvant.
Also disclosed is a vaccine composition comprising: a unit dose form of a prepared peptide comprising an epitope set forth in Table 6, wherein the peptide comprises less than 250, 200, 150, 100, 75, 50, 40, 35, 30, 25, 20 or 15 amino acids that have 100% identity with a native peptide sequence; and, a pharmaceutical excipient. The vaccine composition can comprise less than 250 contiguous amino acids that have 100% identity with a native peptide sequence which is CEA, HER2/neu, MAGE2, MAGE3, or p53. The vaccine composition can comprise at least one epitope selected from Table 6 that is modified by lipidation, i.e., lipidated. The pharmaceutical excipient of the vaccine composition can comprise an adjuvant. A vaccine composition can comprise an antigen presenting cell, wherein at least one epitope selected from Table 6 is bound to an HLA molecule on the antigen presenting cell, whereby a T lymphocyte receptor can bind to a complex of the HLA molecule and the at least one peptide. The antigen presenting cell can be a dendritic cell. The vaccine composition can comprise at least one epitope selected from Table 6 that is comprised by a liposome.
A composition in accordance with any embodiment of the invention can further comprise a PanDR binding molecule, such as aKXVAAWTLKAAa, aKFVAAWTLKAAa, aKYVAAWTLKAAa, aKFVAAYTLKAAa, aKXVAAYTLKAAa, aKYVAAYTLKAAa, aKFVAAHTLKAAa, aKXVAAHTLKAAa, aKYVAAHTLKAAa, aKFVAANTLKAAa, aKXVAANTLKAAa, aKYVAANTLKAAa, AKXVAAWTLKAAA (SEQ ID NO:26), AKFVAAWTLKAAA (SEQ ID NO:27), AKYVAAWTLKAAA (SEQ ID NO:28), AKFVAAYTLKAAA (SEQ ID NO:29), AKXVAAYTLKAAA (SEQ ID NO:30), AKYVAAYTLKAAA (SEQ ID NO:31), AKFVAAHTLKAAA (SEQ ID NO:32), AKXVAAHTLKAAA (SEQ ID NO:33), AKYVAAHTLKAAA (SEQ ID NO:34), AKFVAANTLKAAA (SEQ ID NO:35), AKXVAANTLKAAA (SEQ ID NO:36), or AKYVAANTLKAAA (SEQ ID NO:37) (a=D-alanine, X=cyclohexylalanine). The PanDR molecule(s) can be amnidated.